• Clinical and Experimental Reproductive Medicine
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• 제51권2호
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• pp.158-162
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• 2024

Objective: The aim of this study was to compare the outcomes of in vitro fertilization (IVF) in patients with a poor ovarian response who used methyltestosterone, versus those using a placebo, in an infertility clinic setting. Methods: This clinical trial included 120 women who had undergone IVF with intracytoplasmic sperm injection due to poor ovarian reserve and infertility. The study took place at the Yas Infertility Center in Tehran, Iran, between January 1, 2018 and January 1, 2019. In the intervention group, 25 mg of methyltestosterone was administered daily for 2 months prior to the initiation of assisted reproductive treatment. The control group was given placebo tablets for the same duration before starting their cycle. Each group was randomly assigned 60 patients. All analyses were performed using SPSS ver. 23 (IBM Corp.). Results: The endometrial thickness in the intervention group was 7.57±1.22 mm, whereas in the control group, it was 7.11±1.02 (p=0.028). The gonadotropin number was significantly higher in the control group (64.7±13.48 vs. 57.9±9.25, p=0.001). However, there was no significant difference between the two groups in the antral follicular count. The chemical and clinical pregnancy rates in the intervention group were 18.33% and 15% respectively, compared to 8.33% and 6.67% in the control group. The rate of definitive pregnancy was marginally higher in the intervention group (13.3% vs. 3.3%, p=0.05). Conclusion: The findings of this study suggest that pretreatment with methyltestosterone significantly increases endometrium thickness and is associated with an increase in the definitive pregnancy rate.

• Clinical and Experimental Reproductive Medicine
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• 제42권2호
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• pp.62-66
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• 2015

Objective: To evaluate the effect of a gonadotropin-releasing hormone (GnRH) antagonist protocol using corifollitropin alfa in women undergoing assisted reproduction. Methods: Six hundred and eighty-six in vitro fertilization-embryo transfer (IVF)/ intracytoplasmic sperm injection (ICSI) cycles were analyzed. In 113 cycles, folliculogenesis was induced with corifollitropin alfa and recombinant follicle stimulating hormone (rFSH), and premature luteinizing hormone (LH) surges were prevented with a GnRH antagonist. In the control group (573 cycles), premature LH surges were prevented with GnRH agonist injection from the midluteal phase of the preceding cycle, and ovarian stimulation was started with rFSH. The treatment duration, quality of oocytes and embryos, number of embryo transfer (ET) cancelled cycles, risk of ovarian hyperstimulation syndrome (OHSS), and the chemical pregnancy rate were evaluated in the two ovarian stimulation protocols. Results: There were no significant differences in age and infertility factors between treatment groups. The treatment duration was shorter in the corifollitropin alfa group than in the control group. Although not statistically significant, the mean numbers of matured (86.8% vs. 85.1%) and fertilized oocytes (84.2% vs. 83.1%), good embryos (62.4% vs. 60.3%), and chemical pregnancy rates (47.2% vs. 46.8%) were slightly higher in the corifollitropin alfa group than in the control group. In contrast, rates of ET cancelled cycles and the OHSS risk were slightly lower in the corifollitropin alfa group (6.2% and 2.7%) than in the control group (8.2% and 3.5%), although these differences were also not statistically significant. Conclusion: Although no significant differences were observed, the use of corifollitropin alfa seems to offer some advantages to patients because of its short treatment duration, safety, lower ET cancellation rate and reduced risk of OHSS.

• 한국수정란이식학회지
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• 제24권4호
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• pp.289-292
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• 2009

Mongolia has 80% livestock of total agriculture industry, 170,000 farms are engaged, 2,500,000 of cows that were beef and dairy cows are raised. Despite of Mongolian has great application with milk, there are not clear differences between cow and dairy cattle, and the production of milk is also low. But the milk suppliers are varied (horse, sheep, goat, etc), so that the total milk production is 500 thousand ton per year. It's really considerable to improve the breed of owing to many problems with big differences among milk qualities. For carrying out for first year project, artificial insemination project was operated with 3rd grade Holstein semen that were imported from S. Korea, and initiation and field training were also carried out through appropriate AI technique we developed for Mongolia environment. Local information research and MOU conclusion were done with professor D. Altangerel in May $10^{th}{\sim}13^{th}$, 2009, and development for AI technique and AI equipments were supplied for Mongolia breeding and natural environment in July $10^{th}{\sim}17^{th}$ in 2009. All cows were treated by synchronization for AI. To do this, $PGF_{2\alpha}$ injection were treated for luteal phase cow, if it wouldn't work, try again after 11 days. After confirmation of estrus, AI and AI training were carried out with sperm injection in the uterus or cervix by rectum-vagina method which is common worldwide, the most effective artificial insemination technique. If cows were return to next estrus cycle, second AI was carried out about approximately 21 days after artificial insemination. After 2 months, all cows not showing return estrus should be taken pregnancy test. Every pregnant cow will be cared thoroughly. Total 48 cows administrated by $PGF_{2\alpha}$ for synchronization and after 48 hours 45 cows (93.8%) showing estrus were detected and then artificial inseminate them within who 8 cows (27.8%) showed return estrus. Therefore, Using $PGF_2{\alpha}$ for synchronization is effective to use for Mongolia breeding conditions. There are possibility of base for food production after all, including increase of livestock production in Mongolia by improvement of breeding cow with AI and embryo transfer project.

• 한국가축번식학회지
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• 제21권2호
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• pp.167-176
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• 1997

본 연구는 수정란을 수란우에 이식하기 이전에 형질전환가능 수정란을 선발할 수 있다면 형질전환동물의 생산에 크게 도움이 되므로 3.2 kb $\beta$-actin promoter (lacZ/n대) DNA를 미세주입하여 배반포기배에서의 발현을 확인하여 이들을 선발할 수 있는가를 규명하고자 하엿다. 채란된 난포란은 10%FBS, 5$\mu\textrm{g}$/ml LH, 0.5$\mu\textrm{g}$/ml FSH, 100 unit/ml penicillin 및 100 $\mu\textrm{g}$/ml streptomycin이 함유된 TCM199에 22~24 시간동안 체외성숙을 유도후 5$\mu\textrm{g}$/ml heparin으로 수정능획득을 유도한 1$\times$106 sperm/ml의 정자로 체외수정을 시켰다. 체외수정후 18~20시간째에 vortexing에 의해 과립막세포를 제거하고 원심분리시켜 자/웅전핵이 확인되는 수정란의 핵에 3~4 ng/${\mu}\ell$ lacZ/neo DNA를 미세주입하였다. 모든 수정란의 배양은 3 mg/ml BSA, 20${\mu}\ell$/ml NEM AMINO acids 및 40${\mu}\ell$/ml BME amino acids가 함유되어 있는 CR1aa 배양액에 neo/DNA로 transfected 된 BRL 단층에서 실시하였다. G418에 대한 적정농도를 찾기 위하여 정상적인 수정란에 0, 50, 100 및 200 $\mu\textrm{g}$/ml G418를 첨가하여 배양한 결과 8일째에 30.3%(44/145), 8.7%(13/150), 0.7%(1/151) 및 0% (0/134)의 수정란이 배반포기까지 발달하였다. 그래서 본 실험에서는 일정하게 100$\mu\textrm{g}$/ml G418을 첨가하여 배양하였다. 총 1,127개의 수정란을 미세주입후 G418 없는 배양액에서 710개 (63.0%)가 분할하였다. 미세주입후 48시간째에 2-세포기이상 분할된 수정란을 대조구 및 100$\mu\textrm{g}$/ml G418처리구를 무작위로 할당하여 배양하였으며, 또한 740개의 정상수정란도 같은 반복수로 배양을 실시하였다. 미세주입한 수정란은 8일 후 11.6%(26/255) 및 5.2% (14/267)가 대조구 및 G418 처리구에서 배반포기까지 발달하였으며 정상수정란은 27.2% (151/740)가 배반포기 배까지 발달하였다. 미세주입후 대조구에서는 23.1$\pm$2.6/70.7$\pm$4.7 (32.7%)의 할구가 $\beta$-Gal 활력을 보였고, 반면에 100$\mu\textrm{g}$/ml G418 처리구에서는 40.3$\pm$4.1/48.8$\pm$7.5 (82.6%)가 $\beta$-Gal 활력을 보였다. 비록 mosaic 형태로 외래유전자가 발현되었지만 대조구에서 87.0% (26/30개) 배반포기가 $\beta$-Gal 활력을 보인 반면, G418 처리구에서는 모든 배반포기가 $\beta$-Gal 활력을 보였다 (P<0.05). 그러나 대조구 및 G418 처리구의 ICM colony에서는 영양배엽과 내배엽을 제외한 epiblast에서는 확인되지 않았다. 그러나 이 결과로부터 $\beta$-actin promoter/lacZ gene이 integration되지 않는 것인지 또는 다만 염색 확인이 되지 않는 것인지를 판단할 수는 없다. 이상의 결과는 미세주입후 G418에서 배양한 배반포기배에서는 대부분의 할구에서 주입된 gene을 발현하고 있었으나 ICM colony에서는 특히 epiblast에서는 발현되지 않거나 침묵하고 있었다. 비록 G418 처리구에서 훨씬 더 높은 비율로 주입된 gene이 발현되고 있으나 총세포수는 유의적으로 감소하여 이후 형질전환동물의 생산과 ES like-cell의 설립에는 감소될 것으로 사려된다. 그러나 형질전환 수정란의 선발 및 형질전환동물의 생산능력에 관해서는 더 많은 연구가 필요하다고 사려된다.

• Clinical and Experimental Reproductive Medicine
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• 제26권1호
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• pp.89-96
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• 1999

당뇨병은 생식내분비 조직의 구조나 기능 변화를 유발하여 호르몬 합성 및 분비량의 변화를 야기하고, 정자의 운동성 등에 영향을 미치는 것으로 알려져 왔다. 그러나 부정소와 수정관내 정자의 농도 변화나 수정능력 획득 및 침체반응에 미치는 영향은 잘 알려져 있지 않다. 본 실험에서는 Wistar 쥐에 streptozotocin을 투여하여 당뇨병을 유발시킨 후 3일과 14일에 부정소 각 부위와 수정관내 정자 농도 변화를 조사하고 부정소 꼬리와 수정관내 정자의 침체반응 유도 실험 (acrosome reaction to ionophore challenge test, ARIC test)을 이용하여 정자의 수정 능력을 평가하였다. Streptozotocin을 주사한 후 혈액내 인슐린 및 포도당 농도는 당뇨병 경과 기간이 길어짐에 따라 반비례 관계를 보였다. 부정소 머리와 몸통내 정자의 농도는 3일군에서부터 감소하기 시작하나 꼬리에서는 14일군 $(15.2{\pm}2.1)$에서 대조군 $(28.1{\pm}4.0)$이나 3일군에 $(24.8{\pm}2.9)$비해 유의하게 감소하였다. 수정관내 절자 농도는 14일군이 $0.025{\pm}0.013$으로 대조군과 $(0.108{\pm}0.03)$ 3일군에 $(0.067{\pm}0.046)$ 비해 유의하게 감소하였으며, 3일군도 대조군에 비해 유의한 차이를 보였다. 자발적 첨체반응율은 대조군의 부정소 꼬리정자는 $37.1{\pm}2.4$이고 수정관내 정자는 $49.3{\pm}2.4$로 두 부위간 유의한 차이를 보였다. 3일군과 14일군의 부정소 꼬리와 수정관내 정자의 자발적 첨체반응율은 대조군에 비해 유의하게 증가하였다. 한편 14일군의 수정관내 정자의 자발적 첨체반응율은 대조군이나 3일군에 비해 유의하게 증가하였다. ARIC test 결과 대조군과 3일군에서는 20%이상 차이를 보였으나 14일군에서는 약 8.4% 차이를 보였다. 위의 결과가 부정소의 성숙 조절기능 이상 또는 정자형성 이상에 기인한 것인지는 더 연구되어야 하나 당뇨병 병력이 길어짐에 따라 정자의 수적인 감소, 자발적 침체반응의 증가나 침체반응의 약화가 유발되어 생식능력의 감소 원인으로 작용하는 것으로 사료된다.

• Toxicological Research
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• 제21권2호
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• pp.167-173
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• 2005

Bisphenol A (BPA) is a monomer used in the manufacture of a multitude of chemical products, including epoxy resins and polycarbonate. The objective of this study was to evaluate the effects of BPA administration on reproductive characteristics and blood hematological and chemical values in offspring of pregnant dams treated with BPA. BPA was administrated to pregnant mice by intraperitoneally injection with 0, 0.05, 0.5 and 5.0 mg/kg B.W. for 5 times at 3 days interval on gestation days 1-16. There were no treatment-related effects of BPA on reproductive organ weight in male offsprings at 45 days-of-age, but body weight was the lowest in 5.0mg BPA group when compared to other groups (P<0.05). No differences in semen characteristics (sperm concentration, viability, motility and abnormality) were observed between the control and BPA treatment groups. The WBC, HB, HT, MCV, MCH, MCHC, albumin, BUN and total protein of blood hematological and chemical values in male offsprings were not difference for any treatment groups, but RBC value in BPA groups was significantly increased comparing to the control group (P<0.05). The PLT value was slightly higher in 5.0mg BPA groups than in any other group, but not significantly difference among the experimental groups. In female offsprings, the effects of BPA didn't affect to the body and ovary weight, but the uterus weight in 5.0mg BPA group was slightly heavier than that of control group (P>0.05). No statically significant difference in blood hematological values in female offsprings were observed between the control group and BPA groups, but the concentration of albumin and BUN were significantly higher in 0.5mg BPA group when compared to control and other BPA treatment groups (P<0.05). The histological evaluation of testis and ovary in growing offspring at 45 days-of-age was not difference between the control group and BPA groups, but endometriosis of the uterus in female offspring was dramatically increased in 0.5 and 5.0mg BPA groups. These founding suggest that low concentration of BPA might not have a important role on reproductive ability or blood metabolite in offspring of pregnant dams treated with BPA.

• 한국수정란이식학회지
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• 제27권2호
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• pp.113-119
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• 2012

Epigenetic modification including genome-wide DNA demethylation is essential for normal embryonic development. Insufficient demethylation of somatic cell genome may cause various anomalies and prenatal loss in the development of nuclear transfer embryos. Hence, the source of nuclear donor often affects later development of nuclear transfer (NT) embryos. In this study, appropriateness of porcine embryonic germ (EG) cells as karyoplasts for NT with respect to epigenetic modification was investigated. These cells follow methylation status of primordial germ cells from which they originated, so that they may contain less methylated genome than somatic cells. This may be advantageous to the development of NT embryos commonly known to be highly methylated. The rates of blastocyst development were similar among embryos from EG cell nuclear transfer (EGCNT), somatic cell nuclear transfer (SCNT), and intracytoplasmic sperm injection (ICSI) (16/62, 25.8% vs. 56/274, 20.4% vs. 16/74, 21.6%). Genomic DNA samples from EG cells (n=3), fetal fibroblasts (n=4) and blastocysts from EGCNT (n=8), SCNT (n=14) and ICSI (n=6) were isolated and treated with sodium bisulfite. The satellite region (GenBank Z75640) that involves nine selected CpG sites was amplified by PCR, and the rates of DNA methylation in each site were measured by pyrosequencing technique. The average methylation degrees of CpG sites in EG cells, fetal fibroblasts and blastocysts from EGCNT, SCNT and ICSI were 17.9, 37.7, 4.1, 9.8 and 8.9%, respectively. The genome of porcine EG cells were less methylated than that of somatic cells (p<0.05), and DNA demethylation occurred in embryos from both EGCNT (p<0.05) and SCNT (p<0.01). Interestingly, the degree of DNA methylation in EGCNT embryos was approximately one half of SCNT (p<0.01) and ICSI (p<0.05) embryos, while SCNT and ICSI embryos contained demethylated genome with similar degrees. The present study demonstrates that porcine EG cell nuclear transfer resulted in hypomethylation of DNA in cloned embryos yet leading normal preimplantation development. Further studies are needed to investigate whether such modification affects long-term survival of cloned embryos.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.103-108
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• 1996

Through the previous studies(I,II), it was observed that human follicular fluid(HFF) was more effective than human fetal cord serum(HFCS) on promoting meiotic resumption of oocytes and improving embryonic development of mouse in vitro. On the basis of these results, we have gradually exchanged HFCS with HFF as protein supplement in human ART. This study was performed to investigate the efficiency of HFF on improving the pregnancy rate in ART. Oocytes were retrieved transvaginally from patients treated with pituitary suppression with GnRH-agonist and ovarian stimulation with human menopausal gonadotro-pin(HMG) and pure follicle stimulating hormone(FSH). Aspirated oocytes were rinsed and cultured in TCM-199 containing HFF, and the concentrations of HFF were adjusted to 10, 20, and 30% according to the use for insemination, embryo growth and embryo transfer, respectively. As possible as, we tried to do embryo transfer into fallopian tube to mimic the coincidence of the cell stage with the place of sojourn in vivo, so we performed various ART programs(IVF & ET; in vitro fertilization, ZIFT; zygote intra fallopian-tube transfer, ZIFT & ET) according to the tubal conditions of patients. On the while, intra cytoplasmic sperm injection(ICSI) was used to assist IVF of the patients who had shown poor standard IVF results by immunological or severe male factor. Of the 255 cycles of ART programs using HFF as protein supplement, 118 cycles were turn out to be succeeded in pregnancy(46.2%, per cycle, p<0.05), while 21 pregnancies were achieved in the 69 cycles using HFCS(30.4%). The 255 cycles using HFF were subdivided into cycles with the type of ART programs, and each pregnancy rate of the ART programs were 44.7% (IVF & ET, 76/170 cycles), 53.4%(ZIFT, 31/58 cycles) and 40.7% (ZIFT & ET, 11/27 cycles), respectively. In the 61 ICSI cycles using HFF, 28 cycles succeed in pregnancy(45.9%), while 7 pregnancies were obtained in the 17 ICSI cycles using HFCS. Also the ongoing pregnancy rate in the group using HFF(78.8%, 93/118 cycles) was higher than that in the group using HFCS(61.9%). Therefore, we found that the use of HFF as protein supplement was more suitable and effective than the use of HFCS to improve the pregnancy rate in ART.

• Clinical and Experimental Reproductive Medicine
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• 제31권4호
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• pp.225-234
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• 2004

Objectives: To evaluate the efficacy of GnRH antagonist cetrorelix in women undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) and to determine changes in serum hormone concentrations during cetrorelix administration. Methods: We performed a clinical trial on 30 patients undergoing COH with highly purified follicular stimulating hormone (HP-FSH) and gonadotropin releasing hormone antagonist (GnRHant), cetrorelix. FSH was administrated from day 2 or 3 of cycle with fixed dose and adjusted according to individual response. 0.25 mg of cetrorelix was injected daily subcutaneously from stimulation day 5 until the day of hCG administration. Daily ultrasound monitoring was performed for growing follicles and serum levels of luteinizing hormone (LH), estradiol ($E_2$) and progesterone were measured daily during cetrorelix administration. Up to 4 embryos were transferred. Results: Mean age of enrolled patients was $32.0{\pm}3.4$ years (mean $\pm$ S.D.). All of 30 patients underwent oocyte pick-up, and embryo transfer was done in 28 patients. The total and mean numbers of received oocytes were 196 and $6.5{\pm}4.7$, the number of fertilized eggs was 111, and the fertilization rate was 56.6%. Total duration of FSH administration was $9.2{\pm}2.2$ days and mean of $24.3{\pm}7.7$ ampules of HP-FSH was administered. Total duration of cetrorelix administration was $5.7{\pm}1.9$ days. Serum LH and progesterone levels were maintained in the range of $1.4{\sim}2.9\;mIU/mL$ and $0.3{\sim}0.6\;ng/mL$, which respectively reflected effective prevention of premature LH surge. Clinical pregnancies were achieved in 9 patients, and overall clinical pregnancy rate was 30.0% per oocyte retrieval, and 32.1% per embryo transfer. Conclusion: GnRH antagonist is safe and convenient for COH for IVF-ET and effective with optimal pregnancy rate.

• Environmental Analysis Health and Toxicology
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• 제15권4호
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• pp.123-130
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• 2000

택트 스위치 제조공정 침지액의 주성분인 2-bromopropane독성에 대한 연구로 최단기의 폭로로 농도를 달리하여 3주간 반복 투여 시험을 시행하여, 흰쥐의 혈액 및 세정관의 변화를 관찰하기 위해 투여기간 동안의 체중의 변화, 고환, 간, 신장 등의 장기무게, 혈액화학과 혈액학적 분석 및 고환의 병리조직의 변화 등을 관찰하여 2-bromopropane의 급성투여 조건에서의 중독현상을 비교.분석하였다. 농도를 달리하여 투여에 따른 체중변동은 통계적으로 유의한(P<0.05)체중의 감소를 나타내었다. 1.000 mg/kg 투여군에서 백혈구수, 적혈구수, 혈구용적과 혈색소 농도에서 유의한 변화(p<0.05)를 보였다. 조직병리학적 소견으로 정 소에서는 세정관의 정조세포와 정모세포의 괴사를 볼 수 있었고, 기저막의 비후, 세정관의 Sertoli세포는 광범위하게 세포질성 공포현상을 보여주고 있다. 또한 간질조직에서는 Leydig 세포의 증식을 볼 수 있었다. 2-bromopropane의 손상부위는 조혈과 생식계가 표적으로 생각되며, 고농도 투여가 저농도 투여에 비해 독성이 심하며 독성물질의 양-반응 관계를 보여주고 있다.